Western Blot Analysis of Oxidative Stress Markers

Western blots were performed as previously described[11 (link)]. The following reagents were used: rabbit polyclonal antibody to cyclooxygenase-2 (COX-2) (AbcamInc, Cambridge, MA, United States), glutamate-cysteine ligase modifier subunit (GCLM) (AbcamInc, Cambridge, MA, United States), inducible nitric oxide (NO) synthase (iNOS) (Santa Cruz Biotechnology, Santa Cruz, CA, United States), endothelial NO synthase (eNOS) (Santa Cruz Biotechnology, Santa Cruz, CA, United States), glutamate-cysteine ligase catalytic subunit (GCLC) (AbcamInc, Cambridge, MA), glutathione (GSH) peroxidase (GPx) (AbcamInc, Cambridge, MA, United States), heme oxygenase-1 (HO-1) (AbcamInc, Cambridge, MA, United States), superoxide dismutase (SOD) (Santa Cruz Biotechnology, Santa Cruz, CA, United States), glyceraldehyde-3-phosphate dehydrogenase (GADPH) (Cell Signaling, Danvers, MA, United States), rabbit monoclonal antibody to NF-κB (Cell Signaling, Danvers, MA, United States), catalase (CAT) (Rockland Immunochemicals, Limerick, PA, United States), and mouse monoclonal antibody to nicotinamide adenine dinucleotide phosphate (NAD(P)H) quinone oxidoreductase-1 (NQO-1) (AbcamInc, Cambridge, MA, United States) followed by secondary anti-rabbit or mouse immunoglobulin G (Cell Signaling, Danvers, MA, United States).

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Takasu C., Vaziri N.D., Li S., Robles L., Vo K., Takasu M., Pham C., Farzaneh S.H., Shimada M., Stamos M.J, & Ichii H. (2017). Treatment with dimethyl fumarate ameliorates liver ischemia/reperfusion injury. World Journal of Gastroenterology, 23(25), 4508-4516.

Publication 2017

COX-2) heme oxygenase-1 (HO-1) superoxide dismutase (SOD) glyceraldehyde-3-phosphate dehydrogenase (GADPH) NAD(P)H) quinone oxidoreductase-1 (NQO-1)

Antibody Catalase Cyclooxygenase 2 Endothelial nitric oxide synthase Enos Glutamate cysteine ligase catalytic subunit Glutamate cysteine ligase modifier subunit Glyceraldehyde 3 phosphate dehydrogenase Heme oxygenase 1 Immunoglobulin g Inos Monoclonal antibody Mouse Nf κb Nicotinamide adenine dinucleotide phosphate Peroxidase Quinone oxidoreductase 1 Rabbit Superoxide dismutase Western blots

Corresponding Organization : University of California, Irvine

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Variable analysis

independent variables

Treatments or conditions applied to the samples

dependent variables

Protein expression levels of: cyclooxygenase-2 (COX-2), glutamate-cysteine ligase modifier subunit (GCLM), inducible nitric oxide (NO) synthase (iNOS), endothelial NO synthase (eNOS), glutamate-cysteine ligase catalytic subunit (GCLC), glutathione (GSH) peroxidase (GPx), heme oxygenase-1 (HO-1), superoxide dismutase (SOD), NF-κB, catalase (CAT), nicotinamide adenine dinucleotide phosphate (NAD(P)H) quinone oxidoreductase-1 (NQO-1)

control variables

Experimental procedures and conditions (previously described in reference [11])
Antibodies used for Western blots: rabbit polyclonal antibody to COX-2, GCLM, iNOS, eNOS, GCLC, GPx, HO-1, SOD, rabbit monoclonal antibody to NF-κB, CAT, mouse monoclonal antibody to NQO-1, and secondary anti-rabbit or mouse immunoglobulin G

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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